Animal welfare issues deserve primary consideration when developing an experimental protocol. Researchers have an ethical obligation to minimize pain and distress in laboratory animals. For many years, rabbits, sheep, mice and other species have been used to produce antisera to assorted hormones, viruses, bacteria, etc. These antibodies have contributed in numerous ways to benefit human and animal health. However, to anyone involved in day to day antibody production, it is evident that the risks to the animal of pain and trauma from repeated multiple injections and bleedings and tissue necrosis due to the potent immunostimulants used as adjuvants, are not trivial. Laying hens provide unique advantages for antibody production. They can contribute to decrease animal suffering and distress in 3 ways. First, a single injection of antigen and adjuvant is sufficient to stimulate antibody production for hundreds of days. There is no booster effect in chickens, and therefore there is less animal distress from repeated handling and injections. Second, mg quantities of antibodies are secreted into eggs, thus there is no need to bleed the animal for antibody collection. In some chicken strains egg production proceeds at a rate of 6 eggs/week insuring abundant antibody supplies. Third, newer less toxic adjuvants can potentially be used to elicit a response. Mammalian proteins and peptides are more immunogenic in chickens, presumably due to the wide phylogenetic difference between avies and mammals. This last advantage has yet to be explored. The purpose of the present protocol is to evaluate the effects of a variety of adjuvants and antigen dosage on antibody production in chickens to a relatively small protein. Glucagon was chosen as a protypical immunogen primarily because it is evolutionarily a highly conserved protein which is readily available and relatively cheap. Moreover, because all mammalian glucagon is identical, there are few useful antibodies to glucagon available to the research community. An optimal immunization protocol will be developed by manipulating the type of adjuvant and glucagon dose. The route of administration will be i.m. which is reportedly superior to i.v., s.c. and i.p. in chickens. This pilot project will lay the foundation for determining whether general guidelines can be developed for antibody production in chickens particularly using newer less toxic adjuvants and difficult mammalian antigens.